Electrocompetent Cells
Electrocompetent Cells - Related Videos
07:10
Rapid Protocol for Preparation of Electrocompetent Escherichia coli and Vibrio cholerae
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Cited by 59 •
2013
Electroporation is a commonly employed method for introducing DNA into bacteria in a process known as transformation. Traditional protocols for the preparation of electrocompetent cells are time consuming and labor intensive. This article describes an alternate, rapid, and efficient method for the preparation of electrocompetent cells presently employed by some laboratories.
04:55
Electroporation-Based Delivery of Fluorescent Biomolecules into Bacterial Cells
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2025
Source: Aigrain, L., et al. Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation. J. Vis. Exp. (2015)The video demonstrates the delivery of fluorescent biomolecules into electrocompetent microbial cells using electroporation. The process begins with the introduction of fluorophore-tagged DNA or proteins into a chilled cell suspension. A high-voltage pulse is then applied to create temporary membrane pores, enabling biomolecule entry into the...
10:04
Tractable In Vivo Reprogramming of Tumor Cells to Type 1 Conventional Dendritic Cell-like Cells
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2025
This protocol describes the in vivo reprogramming of mouse cancer cells into type 1 dendritic-like cells within the tumor microenvironment through enforced expression of the transcription factors PU.1, IRF8, and BATF3.
22:06
Derivation of Hematopoietic Stem Cells from Murine Embryonic Stem Cells
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Cited by 8 •
2007
This protocol details the derivation of transplantable hematopoietic stem cells from mouse embryonic stem cells (ESC) and their subsequent injection into lethally irradiated recipient mice. Briefly, ESC are differentiated as embryoid bodies, which are then infected with retroviral HoxB4 and co-cultured with OP9 stromal cells and hematopoietic cytokines.
02:47
Human ES cells: Starting Culture from Frozen Cells
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2006
Here we demonstrate how our lab begins a HuES human embryonic stem cell line culture from a frozen stock.
FAQs
Removing ions before electroporation is essential because the cells must remain in a low-conductivity suspension. The washing step reduces the ionic content surrounding the cells, creating conditions suitable for a brief high-voltage pulse. This preparation connects the physical state of the sample to successful DNA delivery and is therefore a central control point in producing electrocompetent cells.
The electric pulse acts on the cell membrane rather than merely mixing DNA and cells. A brief high-voltage exposure creates transient pores, allowing plasmid DNA or other genetic material to cross the membrane. Because the pores are temporary, the pulse provides a short entry window while allowing the cells to proceed to the recovery stage afterward.
Recovery after the pulse is a distinct stage, not an optional extension of DNA uptake. Cells are transferred into nutrient medium so they can recover after membrane permeabilization. This step follows electroporation and precedes experimental use of the transformed population, linking the physical delivery event to cells that can support cloning or plasmid propagation.
The approach can introduce genetic constructs into bacteria and other cell types, so its relevance is not limited to one microorganism. The same electroporation logic connects membrane permeabilization with delivery of DNA for gene-function studies or engineered biological products, while the selected cell type determines the biological system in which the construct is examined.
Preparation begins by washing the cells to remove ions, then suspending them in a low-conductivity solution. DNA is introduced before the brief high-voltage pulse, and the cells are placed in nutrient medium for recovery afterward. Keeping these stages distinct helps researchers connect sample preparation, DNA entry, and post-pulse recovery in a reproducible workflow.
Successful electroporation depends on a low-conductivity cell suspension and a brief, high-voltage pulse. The low ionic content supports the electrical treatment, while the pulse characteristics produce the transient membrane pores needed for DNA entry. Afterward, nutrient medium supplies the recovery environment. Together, these conditions coordinate DNA uptake and restoration of the cells after pulsing.
Researchers use electrocompetent cells when they need to introduce genetic material for cloning, plasmid propagation, genomic library construction, or recombinant protein research. The method is useful when efficient DNA uptake supports a larger downstream project, such as maintaining genetic constructs, assembling library resources, or investigating the production of engineered biological products.
In biology, the method provides a route from an external genetic construct to an experimental cell system. Introducing DNA into bacterial or other cell types enables studies of gene function and supports research on recombinant proteins and engineered biological products. Thus, the technique links membrane-level events during electroporation with broader questions about how genetic information produces biological outcomes.